Evaluation of MCF10A as a Reliable Model for Normal Human Mammary Epithelial Cells

Abstract

Breast cancer is the most common cancer in women and a leading cause of cancer-related deaths for women worldwide. Various cell models have been developed to study breast cancer tumorigenesis, metastasis, and drug sensitivity. The MCF10A human mammary epithelial cell line is a widely used in vitro model for studying normal breast cell function and transformation. However, there is limited knowledge about whether MCF10A cells reliably represent normal human mammary cells. MCF10A cells were grown in monolayer, suspension (mammosphere culture), three-dimensional (3D) “on-top” Matrigel, 3D “cell-embedded” Matrigel, or mixed Matrigel/collagen I gel. Suspension culture was performed with the MammoCult medium and low-attachment culture plates. Cells grown in 3D culture were fixed and subjected to either immunofluorescence staining or embedding and sectioning followed by immunohistochemistry and immunofluorescence staining. Cells or slides were stained for protein markers commonly used to identify mammary progenitor and epithelial cells. MCF10A cells expressed markers representing luminal, basal, and progenitor phenotypes in two-dimensional (2D) culture. When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability. Cells in mammospheres and 3D culture expressed both luminal and basal markers. Surprisingly, the acinar structure formed by MCF10A cells in 3D culture was positive for both basal markers and the milk proteins β-casein and α-lactalbumin. MCF10A cells exhibit a unique differentiated phenotype in 3D culture which may not exist or be rare in normal human breast tissue. Our results raise a question as to whether the commonly used MCF10A cell line is a suitable model for human mammary cell studies.

GSK-3 inhibition overcomes chemoresistance in human breast cancer

“…2B). MCF10A are non-tumorigenic cells derived from benign proliferative breast tissue that were spontaneously immortalized and are suggested as a benign breast epithelial cell model [28]. We found that pharmacological inhibition of GSK-3 did not significantly affect the growth of MCF10A cells, whereas growth of SKBR3 and various breast cancer cell lines at the same concentrations was suppressed by the GSK-3 inhibitors AR-A014418, 9-ING-41 and 9-ING-87 (Figs.…”

HDAC7‐mediated control of tumour microenvironment maintains proliferative and stemness competence of human mammary epithelial cells

“…Since a recent study claimed a role of HDAC7 in breast cancer stem cells (Witt et al ., ), we compared sphere generation by HDAC7 −/− and HDAC7 +/+ cells. As previously reported (Amin et al ., ; Clément et al ., ; Qu et al ., ), MCF10A can form spheres with low efficiency (Fig. A).…”

Mammalian gene expression variability is explained by underlying cell state

“…To achieve this, we developed an experimental protocol that combines MERFISH, a multiplexed and error robust protocol of counting RNA transcripts using fluorescent in situ hybridization (Chen et al , ; Moffitt et al , ) with rich profiling of the underlying cell state (Fig ). We used the MCF10A mammary epithelial cell line, which is often used in studies of cellular variability due to their nontransformed nature and their accessibility to imaging (Selimkhanov et al , ; Qu et al , ). We focused on genes that share biological function: involvement in the Ca 2+ signaling network, a key pathway important to the cellular response to tissue wounding (Minns & Trinkaus‐Randall, ; Justet et al , ).…”